中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2013年
9期
616-620
,共5页
邱艳%陈洪潭%许国强%虞朝晖%滕晓东%姚洪田
邱豔%陳洪潭%許國彊%虞朝暉%滕曉東%姚洪田
구염%진홍담%허국강%우조휘%등효동%요홍전
窖蛋白1%脂肪肝%高脂血症%胆固醇,膳食%小鼠,近交C57BL
窖蛋白1%脂肪肝%高脂血癥%膽固醇,膳食%小鼠,近交C57BL
교단백1%지방간%고지혈증%담고순,선식%소서,근교C57BL
Caveolin 1%Fatty liver%Hyperlipidemias%Cholesterol,dietary%Mice,inbred C57BL
目的 探索陷窝蛋白-1在高脂饮食性非酒精性脂肪性肝病(NAFLD)发病中的作用.方法 给予12只10周龄雄性C57BL/6小鼠高脂高胆固醇饮食14周,建立NAFLD的动物模型作为实验组.同时,普通饮食饲养同种系小鼠6只作为对照组.第14周末处死全部小鼠,比较两组体质量、肝质量和血脂变化.分别用实时荧光定量PCR(qPCR)和Western印迹法检测陷窝蛋白-1在NAFLD小鼠肝组织中的基因和蛋白表达水平变化.苏木精-伊红染色后光学显微镜下观察小鼠肝脏的脂肪变性情况,免疫组织化学染色后观察陷窝蛋白-1在肝组织中分布的变化.采用t检验比较两组肝组织中陷窝蛋白-1 mRNA及蛋白水平的差异,Mann-whitney U检验分析不同程度脂肪肝小鼠的肝脏陷窝蛋白-1表达的免疫组织化学积分差异.结果 实验组小鼠在高脂高胆固醇饮食14周后全部发生NAFLD,其中重度9只,中度3只.与对照组比较,实验组小鼠的血清TC[(1.940±0.300) mmol/L比(3.771±0.800) mmol/L,t=-3.760]、高密度脂蛋白胆固醇[(1.120±0.066)mmol/L比(2.224±0.420) mmol/L,t=-5.474]、低密度脂蛋白胆固醇[(0.510±0.191) mmol/L比(1.241±0.660) mmol/L,t=-3.332]均显著升高(P均＜0.01),但TG差异无统计学意义(P＞0.05).实验组小鼠肝脏陷窝蛋白[1的mRNA(1.536±0.226比0.980±0.272,t=3.371,P＜0.05)和蛋白水平(0.643±0.240比0.100±0.130,t=-4.847,P＜0.01)均显著高于对照组.免疫组织化学结果显示,表达增加的陷窝蛋白-1主要分布于肝细胞膜、脂滴膜和细胞质中.结论 高脂高胆固醇饮食诱导的NAFLD小鼠肝脏陷窝蛋白1水平上调,可能参与了NAFLD的发病机制.
目的 探索陷窩蛋白-1在高脂飲食性非酒精性脂肪性肝病(NAFLD)髮病中的作用.方法 給予12隻10週齡雄性C57BL/6小鼠高脂高膽固醇飲食14週,建立NAFLD的動物模型作為實驗組.同時,普通飲食飼養同種繫小鼠6隻作為對照組.第14週末處死全部小鼠,比較兩組體質量、肝質量和血脂變化.分彆用實時熒光定量PCR(qPCR)和Western印跡法檢測陷窩蛋白-1在NAFLD小鼠肝組織中的基因和蛋白錶達水平變化.囌木精-伊紅染色後光學顯微鏡下觀察小鼠肝髒的脂肪變性情況,免疫組織化學染色後觀察陷窩蛋白-1在肝組織中分佈的變化.採用t檢驗比較兩組肝組織中陷窩蛋白-1 mRNA及蛋白水平的差異,Mann-whitney U檢驗分析不同程度脂肪肝小鼠的肝髒陷窩蛋白-1錶達的免疫組織化學積分差異.結果 實驗組小鼠在高脂高膽固醇飲食14週後全部髮生NAFLD,其中重度9隻,中度3隻.與對照組比較,實驗組小鼠的血清TC[(1.940±0.300) mmol/L比(3.771±0.800) mmol/L,t=-3.760]、高密度脂蛋白膽固醇[(1.120±0.066)mmol/L比(2.224±0.420) mmol/L,t=-5.474]、低密度脂蛋白膽固醇[(0.510±0.191) mmol/L比(1.241±0.660) mmol/L,t=-3.332]均顯著升高(P均＜0.01),但TG差異無統計學意義(P＞0.05).實驗組小鼠肝髒陷窩蛋白[1的mRNA(1.536±0.226比0.980±0.272,t=3.371,P＜0.05)和蛋白水平(0.643±0.240比0.100±0.130,t=-4.847,P＜0.01)均顯著高于對照組.免疫組織化學結果顯示,錶達增加的陷窩蛋白-1主要分佈于肝細胞膜、脂滴膜和細胞質中.結論 高脂高膽固醇飲食誘導的NAFLD小鼠肝髒陷窩蛋白1水平上調,可能參與瞭NAFLD的髮病機製.
목적 탐색함와단백-1재고지음식성비주정성지방성간병(NAFLD)발병중적작용.방법 급여12지10주령웅성C57BL/6소서고지고담고순음식14주,건립NAFLD적동물모형작위실험조.동시,보통음식사양동충계소서6지작위대조조.제14주말처사전부소서,비교량조체질량、간질량화혈지변화.분별용실시형광정량PCR(qPCR)화Western인적법검측함와단백-1재NAFLD소서간조직중적기인화단백표체수평변화.소목정-이홍염색후광학현미경하관찰소서간장적지방변성정황,면역조직화학염색후관찰함와단백-1재간조직중분포적변화.채용t검험비교량조간조직중함와단백-1 mRNA급단백수평적차이,Mann-whitney U검험분석불동정도지방간소서적간장함와단백-1표체적면역조직화학적분차이.결과 실험조소서재고지고담고순음식14주후전부발생NAFLD,기중중도9지,중도3지.여대조조비교,실험조소서적혈청TC[(1.940±0.300) mmol/L비(3.771±0.800) mmol/L,t=-3.760]、고밀도지단백담고순[(1.120±0.066)mmol/L비(2.224±0.420) mmol/L,t=-5.474]、저밀도지단백담고순[(0.510±0.191) mmol/L비(1.241±0.660) mmol/L,t=-3.332]균현저승고(P균＜0.01),단TG차이무통계학의의(P＞0.05).실험조소서간장함와단백[1적mRNA(1.536±0.226비0.980±0.272,t=3.371,P＜0.05)화단백수평(0.643±0.240비0.100±0.130,t=-4.847,P＜0.01)균현저고우대조조.면역조직화학결과현시,표체증가적함와단백-1주요분포우간세포막、지적막화세포질중.결론 고지고담고순음식유도적NAFLD소서간장함와단백1수평상조,가능삼여료NAFLD적발병궤제.
Objective To explore the role of caveolin-1 in nonalcoholic fatty liver disease (NAFLD) caused by high-fat diet.Methods A total of 12 ten-week-old male C57BL/6 mice were fed with high-fat and high-cholesterol diet for 14 weeks to establish the NAFLD animal model.And six syngeneic mice fed with normal diet at the same time were taken as control.All the mice were sacrificed by the end of 14th week,body weight,liver weight and the changes of serum lipids of the two groups were compared.The changes of caveolin-1 at mRNA and protein levels in the liver of mice with NAFLD were detected by quantitative polymerase chain reaction (qPCR) and Western blot.The liver steatosis of the mice was observed under light microscopy after stained by hematoxylin and eosin.The changes of distribution of caveolin-1 in liver were examined by immunohistochemistry.The differences of caveolin-1 at mRNA and protein level in livers between the two groups were compared by t test.The differences of immunohistochemical scores of caveolin-1 expression in the livers of mice with different degree of fatty liver were analyzed by ordinal variables of two independent samples ranksum test analysis.Results After 14 weeks high-fat and high-cholesterol diet,all the mice of experiment group developed NAFLD.Nine of which were severe and three were moderate.Compared with the control group,serum total cholesterol,high density lipoprotein cholesterol and low density lipoprotein cholesterol of experiment group significantly increased ((1.940 ± 0.300) mmol/L vs (3.771±0.800) mmol/L,(1.120±0.066) mmol/L vs (2.224±0.420) mmol/L,(0.510±0.191) mmol/L vs (1.241±0.660) mmol/L,t=-3.760,-5.474,-3.332,all P＜0.01),however there was no significant difference in triglyceride (P＞0.05).The caveolin-1 of experiment group significantly increased at mRNA (1.536 ±0.226 vs 0.980± 0.272,t=3.371,P＜0.05) and protein levels (0.643±0.240 vs 0.100±0.130,t=4.847,P＜0.01).The immunohistochemical results indicated that the increased caveolin-1 expression mainly distributed in the membrane of hepatocytes,cytoplasm and membrane of lipid droplets.Conclusion The up-regulated caveolin-1 expression in the livers of NAFLD mice induced by high-fat and high-cholesterol may be involved in the mechanism of NAFLD.
